微囊藻毒素LR的定量检测研究  

Study on quantitative determination of microcystins-LR

作　　者：王志慧[1,2] 金玉[3] 张婉静[3] 张爱 黄微 李作生[2,5] 

机构地区：[1]云南农业大学动物科技学院,云南昆明650228 [2]云南沃森生物技术股份有限公司,云南昆明650106 [3]云南省环境监测中心站,云南昆明650034 [4]大理州环境监测站,云南大理671000 [5]成都军区疾病预防控制中心,云南昆明650032

出　　处：《中国卫生检验杂志》2013年第10期2250-2252,共3页Chinese Journal of Health Laboratory Technology

摘　　要：目的制备MC-LR的单克隆抗体,初步建立微囊藻毒素LR定量检测的竞争ELISA方法。方法从云南滇池中收集蓝藻,提取MC-LR精制样品,与牛血清白蛋白偶联,制备成MC-LR-BSA完全抗原,免疫BALB/c小鼠将免疫鼠的脾细胞与SP2/0骨髓瘤细胞融合,制备抗MC-LR特异性单克隆抗体。应用制备的单克隆抗体对不同稀释浓度的MC-LR样品进行竞争ELISA定量检测。结果制备了3株抗MCLR的单克隆抗体,MC-LR样品的竞争ELISA定量检测显示浓度从高到低的相应A值呈现明显的递增梯度趋势。结论制备的MC-LR单克隆抗体能够用于MC-LR的定量检测。Objective To prepare the monoclonal antibody against MC - LR and to establish the competitive inhibi- tion enzyme linked immunosorbent assay （ciELISA） method for quantitative determination of MC - LR. Methods The refined MC - LR was extracted from blue - green algae in the Dian Lake in Yunnan province and prepared to be MC - LR - BSA complete antigen by coupling with Bovine serum albumin. Then Balb/c mice were immunized the complete antigen. The splenocyte from the immunized mice was fused with SP＇2/0 myeloma cell to prepare the MC - LR specific monoclonal antibody for quantitative determination of MC - LR samples by competitive inhibition enzyme linked immunosorbent assay. Results Three hybridoma secreting monoclonal antibodies（McAb） against MC -LR were achieved by fusion of both spleen cells of Balb/c mice immunized with MC - LR - BSA and SP2/0 myeloma cells. The increasing tendency of OD with ciELISA presented corresponding concentration of MC - LR sample from high to low. Conclusion MC - LR. The monoclonal antibody prepared against MC - LR can be used to detect the content of MC - LR.

关 键 词：微囊藻毒素 完全抗原 单克隆抗体 竞争酶联免疫吸附试验 

分 类 号：O657.63[理学—分析化学]